Enhanced Therapeutic and Long-Term Dynamic Vascularization Effects of Human Pluripotent Stem Cell-Derived Endothelial Cells Encapsulated in a Nanomatrix Gel

• Type of Material: Article
• Authors: Shin-Jeong Lee, Emory University; Young Doug Sohn, Emory University; Adinarayana Andukuri, Emory University; Sangsung Kim, Emory University; Jaemin Byun, Emory University; Ji Woong Han, Emory University; In-Hyun Park, Yale University; Ho-Wook Jun, University of Alabama Birmingham; Young-sup Yoon, Emory University
• Language: English
• Date: 2017-11-14
• Publisher: American Heart Association
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2017 American Heart Association, Inc.
• Final Published Version (URL): http://dx.doi.org/10.1161/CIRCULATIONAHA.116.026329
• Title of Journal or Parent Work: Circulation
• ISSN: 0009-7322
• Volume: 136
• Issue: 20
• Start Page: 1939
• End Page: +
• Grant/Funding Information: This work was supported by grants from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (No HI16C2211 and HI15C2782), the Bio & Medical Technology Development Program of the National Research Foundation (NRF) funded by the Korean government (No 2015M3A9C6031514 and 2016R1D1A1B03933154), National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) (No DP3-DK094346 and DP3-DK108245), National Heart, Lung, and Blood Institute (NHLBI) (No R01HL127759 and R01HL129511), and NIH 1R01HL125391-01 to Y-s.Y. and H-W. J.
• Keywords: CD31(+) CELLS; GENERATION; Life Sciences & Biomedicine; stem cells; Cardiovascular System & Cardiology; TRANSPLANTATION; pluripotent stem cells; IN-VITRO; CARDIOMYOCYTES; vascular diseases; BONE-MARROW; DIFFERENTIATION; Cardiac & Cardiovascular Systems; regeneration; endothelial cells; Peripheral Vascular Disease; Science & Technology; MOUSE MODEL; DISEASE; MYOCARDIAL-INFARCTION
• Research Categories: Health Sciences, General

Abstract

BACKGROUND: Human pluripotent stem cell (hPSC)-derived endothelial cells (ECs) have limited clinical utility because of undefined components in the differentiation system and poor cell survival in vivo. Here, we aimed to develop a fully defined and clinically compatible system to differentiate hPSCs into ECs. Furthermore, we aimed to enhance cell survival, vessel formation, and therapeutic potential by encapsulating hPSC-ECs with a peptide amphiphile (PA) nanomatrix gel. METHODS: We induced differentiation of hPSCs into the mesodermal lineage by culturing on collagen-coated plates with a glycogen synthase kinase 3β inhibitor. Next, vascular endothelial growth factor, endothelial growth factor, and basic fibroblast growth factor were added for endothelial lineage differentiation, followed by sorting for CDH5 (VE-cadherin). We constructed an extracellular matrix-mimicking PA nanomatrix gel (PA-RGDS) by incorporating the cell adhesive ligand Arg-Gly-Asp-Ser (RGDS) and a matrix metalloproteinase-2-degradable sequence. We then evaluated whether the encapsulation of hPSC-CDH5+ cells in PA-RGDS could enhance long-term cell survival and vascular regenerative effects in a hind-limb ischemia model with laser Doppler perfusion imaging, bioluminescence imaging, real-time reverse transcription-polymerase chain reaction, and histological analysis. RESULTS: The resultant hPSC-derived CDH5+ cells (hPSC-ECs) showed highly enriched and genuine EC characteristics and proangiogenic activities. When injected into ischemic hind limbs, hPSC-ECs showed better perfusion recovery and higher vessel-forming capacity compared with media-, PA-RGDS-, or human umbilical vein EC-injected groups. However, the group receiving the PARGDS-encapsulated hPSC-ECs showed better perfusion recovery, more robust and longer cell survival (> 10 months), and higher and prolonged angiogenic and vascular incorporation capabilities than the bare hPSC-EC-injected group. Surprisingly, the engrafted hPSC-ECs demonstrated previously unknown sustained and dynamic vessel-forming behavior: initial perivascular concentration, a guiding role for new vessel formation, and progressive incorporation into the vessels over 10 months. CONCLUSIONS: We generated highly enriched hPSC-ECs via a clinically compatible system. Furthermore, this study demonstrated that a biocompatible PA-RGDS nanomatrix gel substantially improved long-term survival of hPSC-ECs in an ischemic environment and improved neovascularization effects of hPSC-ECs via prolonged and unique angiogenic and vessel-forming properties. This PA-RGDS-mediated transplantation of hPSC-ECs can serve as a novel platform for cell-based therapy and investigation of long-term behavior of hPSC-ECs.